| # | First_name | Last_name |
|---|---|---|
| 1 | Julia | Alten |
| 2 | Monika | Brüggemann |
| 3 | Mireia | Camós |
| 4 | Gianni | Cazzaniga |
| 5 | Emmanuelle | Clappier |
| 6 | Conny | Eckert |
| 7 | Sarah | Elitzur |
| 8 | Nicola | Gökbuget |
| 9 | Csongor | Kiss |
| 10 | Josep Maria | Ribera |
| 11 | Nick | Short |
| 12 | Dave | Teachey |
| 13 | Jan | Trka |
| 14 | Bela | Wrench |
| 15 | Vincent | van der Velden |
first-line: AIEOP BFM ALL 2017, ALLTogether 1 second line: IntReALL 2010 HR, ALL-REZ BFM Registry, soon IntReALL BCP 2920, HemiSmart
We only work with the second line therapy protocols.| Category | Comment |
|---|---|
| Other | It is recommended that immunophenotyping is performed in a central lab, but it is only prtially done centrally |
| Other | All samples are analyzed by accredited, “matured” national central laboratories. |
| Other | Diagnostic flow is centralized in 11 reference laboratories |
| Category | Comment |
|---|---|
| Other | MRD Flow is only done if no molecular MRD marker is available |
| Other | All centralized except Monza center. |
| Other | See reply to previous Q. |
| Other | Flow-MRD is centralized in 11 reference laboratories |
| Other | B-ALL on trial: COG approved labs. Off study local labs. Infant and T-ALL are central labs. |
| Category | Comment |
|---|---|
| Other | Ig/TC central, BCR::ABL partly central |
| Other | All centralized except Padua center |
| Other | Molecular MRD is being performed as reasearch projects by some ALLIC national team laboratories. The method is not yet available for diagnostic purposes. |
| Other | Molecular-MRD is centralized in 2 accredited laboratories at H. Sant Joan de Deu (Barcelona) and H. Niño Jesús (Madrid) |
| Other | Molecular MRD is performed in subsets of patients with B-ALL |
| Category | Comment |
|---|---|
| Other | partly overlapping |
| Other | Flow-MRD and molecular-MRD are performed in the same lab only in the 2 molecular reference labs. For the rest of patients, samples are sent for the corresponding Flow-MRD lab and one of the two accredited molecular-MRD laboratories |
| Category | Comment |
|---|---|
| Other | several vials are taken. In theory the first sample should be send to molecular MRD (except day 15) and the second/third sample to Flow Cytometry and Moelcular genetics. |
| Category | Comment |
|---|---|
| Other | Not systematically performed |
| Other | before and after allo-SCT, after relapse, after CART |
| Other | We follow the protocol. Additionally, in case of relapse suspicion based on clinical or laboratory findings |
| Other | Before and after allo-SCT |
| Other | all because all are relapses |
| Category | Comment |
|---|---|
| Other | depends on a situation, can be more than four times, follow up post-SCT is usually day30, day 60, day 180, 1 yr, 2yr, 3yr |
| Other | Never. Transplant is not included on our trials. |
| Other | currentl not defined, very frequent |
| Other | within the first year after each block |
| Other | depending of the MRD-kinetics and the risk profile (3-5 x in HR without alloHSCT and 8-10 x in pts with alloHSCT) |
| Category | Comment |
|---|---|
| Clinical Signs | prolonged cytopenias |
| Clinical Signs | Standard risk (after TP1)/Medium risk patients (after TP2): Only if there are clincal or laboratory signs of relapse (e.g. prolonged cytopenia…) |
| Fixed Intervals | Every 3-4 months |
| Fixed Intervals | evry three months at 1st year, every six months for the 2nd year and then yearly up to 5 years |
| Fixed Intervals | Ph negative ALL, chemotherapy: first year: prior to each consolidation cycle, second+third year: every 3 months, fourth year: evey 6 months. Ph negative ALL, after SCT: every 3 months. Ph+ ALL, after SCT: every 6 weeks in blood, every 3 months in BM. |
| Fixed Intervals | every 3 months (three years)/ 2 months (2 years), and if there are clinical signs of relapse |
| Fixed Intervals | 3 |
| Fixed Intervals | depends on a situation, follow up post-SCT is usually day 30, day 60, day 180, 1 yr, 2yr, 3yr, in relapsed based on relapse protocol |
| Fixed Intervals | end of therapy |
| Fixed Intervals | MRD post alloHSCT (d+30, d+60, d+100, d+180, d+365) |
| Other | If there are clinical/lab signs suspicious for relapse, and in pts. after alloHSCT |
| Other | after each treatment block, ad hoc practice in maintenance every 3 months |
| Other | MRD after each element of HR-Therapy (AIEOP-BFM:all HR-patients: TPHR1, TPHR2, TPHR3 (=MRD after each HR-Block), additionally all HR-T-ALL, all Infants, B-ALL (not atl east 2 x negative in subsequent time points): prior to Re-Induction (prior to 2. Protocol III and 3. Protocol III) |
| Category | Comment |
|---|---|
| Dx Months | ~36 months |
| Dx Months | until end of year 4 (chemotherapy patients) |
| Dx Months | uptill 5 years after reaching CR |
| Dx Months | approximately 36-48 months |
| Dx Months | x1 at end of therapy |
| Dx Months | end of intensive chemotherapy (see question 16) |
| Transplant Months | Not fully standardized. Depends on MRD kinetics. |
| Transplant Months | approximately 36 months |
| Transplant Months | see above, in negative samples post-SCT last BM is 3 years post-SCT, after that yearly PB. But depends on the clinical situation and individual decision of physicians |
| Transplant Months | 1 year |
| Lifelong | evry three months at 1st year, every six months for the 2nd year and then yearly up to 5 years |
| Other | evry three months at 1st year, every six months for the 2nd year and then yearly up to 5 years |
| Response | Answer |
|---|---|
| Other | Positivity below QR: warning signal, MRD >1XE-04: molecular relapse |
| Other | 5x10-2 |
| Other | any MRD positivity, but in positive not quantifiable verified by NGS |
| Other | Any MRD-positivity with further confirmation with other method and different sample |
| Other | We follow PDL guidelines |
| Other | For relapse definition more than 1 method is needed for confirmation, if MRD is < 5%. MRD of 5E-04 is highly suspicious of pending relapse if detected late in therapy |
| Response | Answer |
|---|---|
| Other | not fully standardized, probably like in Ph-negative ALL. |
| Other | 5x10-2 |
| Other | we do BCR/ABL on DNA level simultaneously and solve individually (CML-like cases) |
| Other | Any MRD-positivity with further confirmation with other method and different sample |
| Other | We follow PDL guidelines |
| Other | do not use IG/TR-MRD; 1x10-2 for flow |
| Other | see question 18 |
| Response | Answer |
|---|---|
| Other | Likely not relevant with IG/TR testing |
| Other | none |
| Other | reported in the context of simultaneous measurement of DNA-based BCR/ABL and Ig/TR (e.g. BCR/ABL 1E-02 and Ig/TR negative does not need to be relapse in CML-like cases) |
| Other | ALLIC group did not define MRD cut-off value for diagnosing molecular relapse in Ph+ALL. LOQ in CML is 1e-05 |
| Other | Any MRD-positivity with further confirmation with other method and different sample |
| Other | We follow PDL guidelines |
| Other | BCR::ABL1 positivity without Ig/TCR positivity and zytomorphological absence of blasts is not considered a relapse (this constellation is suspicious for misdiagnosis in the beginning –> CML in blasts crisis ) |
| Category | Comment |
|---|---|
| Yes, always | Yes if converts from negative to positive but only if there is high confidence that the finding reflects true MRD (e.g. not “background”), ideally confirmed on a follow-up sample. |
| Dependes on MRD level | If ther is higher than 1x10e-4, confirmed in a second sample |
| Dependes on MRD level | 1xE-04 |
| Dependes on MRD level | if >0,01% |
| Dependes on MRD level | 10-4 but this is defined then as molecular relapse which is different from hematologic relapse |
| Dependes on MRD level | Clearly if 1E-02, but it would trigger clinical actions if 1E-04 if confirmed by 2 tecniques in 2 different consecutive samples |
| Dependes on MRD level | 1x10-2 |
| Dependes on MRD level | one log increase required |
| Dependes on MRD level | MRD 1 -<5%: two other tests with >=1% or second BM. MRD <25% one additional test >= 1%. doi: 10.1182/blood.2021012328. PMID: 34192312; PMCID: PMC8952186. |
| No, never | depends on ALL type-different situation in CML-like ALL, see above. In Ph-neg ALL pos NQ after negative always verified by NGS |
| No, never | Only if MRD exceedes the 1% (1–02) limit for CR as defined by the P-d-L paper |
| Depends on additional findings | MRD-positivity conversion should be unambiguous: either high level, or if low, without any doubt (confirmed on a second sample or a second marker or technique, or with clinical signs of extramedullary relapse)) |
| Depends on additional findings | Flow or/and morphology |
| Response | Answer |
|---|---|
| Other | depends on the context and MRD level |
| Other | We follow PdL so depends on if a confirmatory test and level of detection |
| Other | depends on the level <10-4 likely requires repeat |
| Other | No, a single positive result is sufficient: depends on the Level (see 21) |
| Response | Answer |
|---|---|
| Other | Only if accompanied by MRD by IG/TR |
| Other | IG/TR to be analyzed |
| Other | Relapse should be confirmed by IG/TR MRD in bone marrow |
| Other | in CML like completely different situation, usually repeated sample necessary |
| Other | We follow PdL |
| Other | no routine BCR::ABL1 monitoring performed |
| Category | Comment |
|---|---|
| Other | No clear rule |
| Other | 2 weeks |
| Other | We follow PdL |
| Other | <1 week for positive >1% without morphology, 2-4 weeks for lower level |
| Response | Answer |
|---|---|
| Other | it depends on MRD levels, frequently, a further BM is planned to monitor increase |
| Other | No defined rule, to be discussed |
| Other | depends on the protocol, usually SCT with salvage before |
| Other | It depends on the MRD level, time point and the clinical setting |
| Other | If meets PdL criteria for relapse we treat as a relapse |
| Other | answers depend of the level of MRD positivity: >=5E-04 after TP2 (day 78/92) Indication for alloHSCT and intensification of therapy. Below 5E-4: Obersevation without immediate intervention |
| Response | Answer |
|---|---|
| Other | depending on CR time, Relapse within 1 year: salvage; if CR>1 year first line induction rretarted |
| Other | A further BM is planned to monitor increase |
| Other | No defined rule, to be discussed |
| Other | not measured in non-HR patients, in HR patients is defined by protocol |
| Other | It depends on the MRD level, time point and the clinical setting |
| Other | If meets PdL criteria for relapse we treat as a relapse |
| Other | it depends on the MRD level see question 25 |
| Response | Answer |
|---|---|
| Other | not fully standardized |
| Other | if no GVHD: reduce immune suppression. Otherwise DLI or blina |
| Other | reduction of immunosuppression; short term confirmation of MRD increase on another BM sample |
| Other | based on the level-low MRD IST tapering, DLI, in high level CART, 2nd SCT.. |
| Other | Individual decision made by the transplant team |
| Other | It depends on the MRD level, time point and the clinical setting |
| Other | Depends on type of transplant, availability of DLI, ability to add a targeted or immunotherapeutic agent |
| Other | it depends on the MRD level. Low positivity: reduction of Immunosuppression if possible, DLI |
| Category | Comment |
|---|---|
| Other | There is a manual that states: 4.6.3 Material 10 ml Knochenmark und/oder 10 ml peripheres Blut Von jedem Untersuchungszeitpunkt, möglichst 1. Aspirat EDTA bevorzugt, Heparin auch möglich Die Zeitpunkte der Remissionskontrollen und Einsendung von Probenmaterial zur MRD-Untersuchung sind den aktuellen Protokollen bzw. Therapieempfehlungen zu entnehmen. Praktisch wird die KM-Punktion jeweils nach Regeneration durchgeführt. Es handelt sich also nicht um Aplasie-Punktionen. In addition, we hav a local SOP in Kiel |
| Other | instructions are give in an extra document of the sample request form |
| Response | Answer |
|---|---|
| Other | research on add-on projects |
| Other | For pilot research purposes |
| Other | molecular genetics |
| Response | Answer |
|---|---|
| Other | See Kiel SOP |
| Other | Don’t know if it is standardized |
| Other | do not know and I am afraid to go ask somewhere because it seems impossible to save the answers and I would loose too much work |
| Other | systems differ between clinical departments |
| Response | Answer |
|---|---|
| Other | not applicable |
| Other | 2-5 ml left and 2-5 ml right |
| Other | depending of age and collection system all of the answers above |
| Response | Answer |
|---|---|
| Other | in multiple aspirates |
| Other | Following the SOP , we perform 2 aspirates of 4ml (1st aspirate) and 10ml (2nd aspirate at different angle through same skin puncture) |
| Other | we do repeat pulls of approximately 2ccs with reanchoring |
| Other | depending of age and collection system all of the answers above |
| Category | Comment |
|---|---|
| Other | we receive DNA already extracted |
| Other | Optimally 2.0 mL |
| Other | depends on the cell number |
| 6–10 mL | depending of age and collection system all of the answers above |
| Category | Comment |
|---|---|
| No, never | We send the sample to the central lab for FCM and there check for hemodilution |
| Yes, using Cytomorphology | Presence/absence of bone marrow particles |
| Yes, using Cytomorphology | assessment of spicules |
| Yes, using Cytomorphology | Important for remission evaluation at day 33 (TP1) |
| Yes, using Flow cytometry | checked for but no calculation, remark made in conclusion |
| Yes, using Flow cytometry | do not know and do not want to go ask FC guys because it seems impossible to save the answers in the process. can send later |
| Yes, using Flow cytometry | Presence of <2% of EBL |
| Yes, using Flow cytometry | Only day 15: Important for FCM-MRD evaluation |
| Category | Comment |
|---|---|
| Other | yes, when blasts are below 50% |
| Other | yes if it is below 50% |
| Other | this varies based on each center practice |
| Other | we adjust the standard curve according to cytomorphology, if blast infiltration is below 50% however we are trying to implement Flow after Ficoll at the moment in order to be more exact |
| Category | Comment |
|---|---|
| Other | I do not know |
| Other | and after for defining blast percentage for molecular standard curve |
| Other | before, in some situations (low number of blasts, sorting for RNAseq) later |
| Other | The complete Flow cytometry analysis is performed before Ficoll separation. After Ficoll separation we perform an additional tube to calculate the blast percentage post-Ficoll |
| Other | planned: after ficoll |
| Category | Comment |
|---|---|
| Other | not performed |
| Other | cytomorphology is performed on BM smears and not in the same sample |
| Category | Comment |
|---|---|
| Other | depends on target, number of classical targets |
| Other | no, not at the moment |
| Other | If flow unavailable |
| Other | SIL-TAL and IKZf1 |
| Category | Comment |
|---|---|
| NA | I do not know |
| Only at certain time points (please specify) | Not in aplasia |
| Only at certain time points (please specify) | in case of low cellularity |
| NA | Sometimes |
| Only at certain time points (please specify) | only remission samples (day 33 onwards), not day 15 and not inital diagnosis |
| Category | Comment |
|---|---|
| Other | I do not know ( I am clinician) |
| Other | Depends on the time-point. In ALL-BFM, we do day 15 MRD, and there we acquire 500,000 cells. In GMALL and in ALL-BFM at other time-points, we aim for 4,000,000 events. |
| Other | Dx 100,000, day 15 100,000-500,000, day 33 more than 1,000,000 |
| Other | 500,000 cells |
| Other | Varies based on lab and method |
| Other | remission spamples:1-10. Mio, Initial + day 15 : 100.000-500,000 |
| Category | Comment |
|---|---|
| Other | do not know ( I am clinician) |
| Other | 10 events |
| Other | Varies based on lab and method |
| Category | Comment |
|---|---|
| NA | do not know ( I am clinician) |
| Other | 40 events |
| Other | Varies based on lab and method |
| Response | Answer |
|---|---|
| Other | BCP-ALL: E-05, T-ALL: rather E-04 or less |
| Other | depends on timepoint, see above |
| Other | 2X10-5 |
| Other | It varies according to the time point |
| Other | day 15: 1E-03 (without bulk lyisis) |
| Response | Answer |
|---|---|
| Other | BCP-ALL: 5xE-05, T-ALL: rather E-04 or less |
| Other | day 15 0.1%, day 33 0.1%-0.01% |
| Other | 6X10-5 |
| Other | It varies according to the time point |
| Category | Comment |
|---|---|
| Commercial | NSG MRD is commercial |
| Commercial | ClonoSeq for NGS-based |
| EuroFlow | BCP-ALL: EuroFlow, but as in-house assay. T-ALL: in-house assay according to BFM-Flow SOP |
| EuroFlow | BCP-ALL MRD assay |
| EuroFlow | BD CytognosTM B-cell Precursors ALL MRD |
| EuroFlow | B-MRD plus 1 additional in-house tube |
| In-house | I-BFM standard |
| In-house | Flow is both performed locally and centrally and depends on population and trial and timepoints |
| In-house | St. Jude in house |
| In-house | T-MRD |
## No comments available.
## No comments available.
## No comments available.
## No comments available.
| Category | Comment |
|---|---|
| Other | This is centrally done and we are not privy to the amount needed |
| Category | Comment |
|---|---|
| Other | This is proprietary and we are not provided it |
| Category | Comment |
|---|---|
| Commercial | clonoSEQ |
| Commercial | Clonoseq |
## No comments available.
| Category | Comment |
|---|---|
| In-house | RT-PCR; note, we use RNA rather than DNA |
| In-house | patient-specific assays |
no responses
| Category | Comment |
|---|---|
| Other | depends on situation (we use GUSB) |
| NA | not applicable for DNA-based analysis |
| Category | Comment |
|---|---|
| NA | not applicable for DNA-based analysis |
| Category | Comment |
|---|---|
| NA | not applicable for DNA-based analysis |
| Category | Comment |
|---|---|
| NA | not applicable for DNA-based analysis |
| Category | Comment |
|---|---|
| In-house | patient-specific assay |
| Category | Comment |
|---|---|
| In-house | patient-specific assay |
| Category | Comment |
|---|---|
| NA | do not know ( I am clinician) |
| Other | aim for >100,000 ABL1 copies |
| Other | NA (Cepheid) |
| Other | depends on situation, in poor samples we use bad quality samples and comment |
| Other | 15-35 copies GAPDH |
| Category | Comment |
|---|---|
| NA | do not know ( I am clinician) |
| Other | cepheid |
| Category | Comment |
|---|---|
| NA | do not know ( I am clinician) |
| Category | Comment |
|---|---|
| NA | do not know ( I am clinician) |
| Other | Only if a clinical trial asks us to do so |
| Category | Comment |
|---|---|
| Commercial | Cepheid |
| Commercial | Qiagene |
| Commercial | Xpert BCR-ABL ULTRA (Cepheid) |
| Commercial | commercial for CML |
| In-house | IRMM ERM-AD623 Plasmid and in-house assay |
| In-house | in house for MRD in Ph+ ALL |
| Category | Comment |
|---|---|
| Commercial | Ipsogen BCR::ABL1 m-bcr Kit |
| Commercial | Cepheid |
| Commercial | Qiagene |
| Commercial | commercial for CML |
| In-house | in house for MRD in Ph+ ALL |
no responses
no responses
## No comments available.
## No comments available.
no responses
| Category | Comment |
|---|---|
| Other | do not use DNA for BCR-ABL |
| Other | only IG/TR performed |
| Other | We do not perform BCR::ABL1-based DNA MRD. However, in case such result was available, both results would be reported from the lab to the clinicians. The concordance or discordance of results and final conclusions are discussed in a multidisciplinary meeting for every MRD assessment for all patients |
| Other | do not use IG/TR-MRD |
| Other | BCR::ABL1 is not used for stratification |
| Category | Comment |
|---|---|
| Other | Both results are reported from the lab to the clinicians. The concordance or discordance of results and final conclusions are discussed in a multidisciplinary meeting for every MRD assessment for all patients. However, in the EsPhALL trial only the IG-TR MRD results are considered. |
| Other | do not use IG/TR-MRD |
| IG/TR-based MRD result | BCR::ABL1 is not used for stratification |
| Category | Comment |
|---|---|
| Other | We do not perform KMT2A-based MRD. However, in case such result was available, both results would be reported from the lab to the clinicians. The concordance or discordance of results and final conclusions are discussed in a multidisciplinary meeting for every MRD assessment for all patients. Generally speaking, we would consider the higher MRD value |
| Other | do not use IG/TR-MRD |
| Category | Comment |
|---|---|
| Other | both results with comment from FC group |
| Other | Both results are reported from the lab to the clinicians. The concordance or discordance of results and final conclusions are discussed in a multidisciplinary meeting for every MRD assessment for all patients. In switch ALL cases, we usually find higher MRD values with IG/TR and consider this higher MRD result for taking clinical actions. |
| Other | in the US all results have to be reported if a clinical test. Protocols can give guidance and local institutions have their own standard. But it is not allowed for the lab who performs the assay to prioritize |
MRD and cytomorphology results are not always concordant. In the following situations, which value do you choose for clinical reporting to state the blast percentage at the time of remission assessment?
MRD and cytomorphology results are not always concordant. In the following situations, which value do you choose for clinical reporting to state the blast percentage at the time of relapse assessment?
| Category | Comment |
|---|---|
| Other | we sometimes receive CSF, not systematically recommended |
| Other | if asked and sent by clinicians |
| Other | rarely to confirm antigen expression following identified CNS involvement |
| Other | only in case of suspected relapse |
| Category | Comment |
|---|---|
| Other | only CNS3 |
| Other | No, but performing flow-cytometry assessment is strongly recommended by the protocol to confirm the presence of blasts identified by cytomorphology |
| Other | not sure, not standardized |
| Category | Comment |
|---|---|
| Other | we sometimes receive SCF, but it is not systematically recommended |
| Other | if asked and sent by clinicians |
| Other | not sure, not standardized |
| Other | rarely to confirm antigen expression following identified CNS involvement |
| Other | only in case of suspected CNS-relapse/persisting blasts in cytospin |
| Category | Comment |
|---|---|
| Other | if MRD+ switch of therapy |
| Other | Each assessment is discussed in a joint meeting of the laboratories and the clinical teams |
| Category | Comment |
|---|---|
| Other (please comment) | We do not use IG/TR qPCR |
| Other (please comment) | We do not perform IG/TR MRD in CSF |
| According to CSF-specific rules (please comment) | not standardized yet |
| Category | Comment |
|---|---|
| NA | do not know ( I am clinician) |
| Category | Comment |
|---|---|
| No | I cannot successfully answer to many technical comments, as I am a clinician |
| Yes | some questions are difficult to answer, this answers would be much more complicated. |
| No | level of control gene in DNA-based methods (which is acceptable)-there are transcript copies. |
| Yes | Communication between the different labs and between the lab and the clinical teams is crucial to discuss both the concordant and the discordant results. We perform an integrated biological report at diagnosis and at each MRD time point, and each BM assessment is discussed weekly with the clinicians. |
A possible topic to discuss is the presence of integrated reports and the (fluid)communication between clinicians and laboratory staff.
Comment for questions 148-153: both the cytomorphology and the MRD results are reported from the lab, and any discordances are studied and discussed in a meeting with the lab and clinical teams to try to explain the discordance (hemodilution of the sample, regenerative BM with a high percentage of B-cell normal precursors (hematogonies), difficult LAIPs, etc) and agree on a conclusion on the results. | |Yes |planned / expected changed in the near future | |Yes |diagnostic procedures and laboratory practices will be country specific, in the UK we operate via the SIHMDS model. For instance, it is not our practice to regress or censor multimodal results for remission to one output, all our reported out to the requesting clinician. In addition practices are guided by the treatment approval criteria - e.g. flow based assessments for CAR-T indication but molecular MRD (plus flow) for Blina for instance, so no singular system exists addressing all the clinically actionable outcomes of MRD testing |